2017
DOI: 10.1186/s13068-017-0732-1
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Molecular and biochemical characterization of recombinant cel12B, cel8C, and peh28 overexpressed in Escherichia coli and their potential in biofuel production

Abstract: BackgroundThe high crystallinity of cellulosic biomass myofibrils as well as the complexity of their intermolecular structure is a significant impediment for biofuel production. Cloning of celB-, celC-encoded cellulases (cel12B and cel8C) and peh-encoded polygalacturonase (peh28) from Pectobacterium carotovorum subsp. carotovorum (Pcc) was carried out in our previous study using Escherichia coli as a host vector. The current study partially characterizes the enzymes’ molecular structures as well as their catal… Show more

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Cited by 12 publications
(18 citation statements)
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“…CelC307 showed the highest cellulolytic activity against soluble CMC substrate (β-1 → 4 linkage) used to calculate kinetic parameters. Compared to some other studied cellulases 19 , 52 55 , CelC307 showed low K m and high k cat /K m , indicating a high preference of the enzyme for CMC that led the enzyme to hydrolyze β- 1 → 4 linkages more rapidly than other evaluated cellulases.…”
Section: Discussionmentioning
confidence: 72%
“…CelC307 showed the highest cellulolytic activity against soluble CMC substrate (β-1 → 4 linkage) used to calculate kinetic parameters. Compared to some other studied cellulases 19 , 52 55 , CelC307 showed low K m and high k cat /K m , indicating a high preference of the enzyme for CMC that led the enzyme to hydrolyze β- 1 → 4 linkages more rapidly than other evaluated cellulases.…”
Section: Discussionmentioning
confidence: 72%
“…However, most of the presently used endoglucanases, especially fungal endoglucanase, usually displayed their highest activity at acidic condition. Their activities are greatly reduced in case of the pH value above 7.0, limiting the application of those fungal enzymes under neutral and alkaline condition (Wang et al, 2014;Zeng et al, 2016;Ibrahim et al, 2017). Therefore, there has a large market and great demand for the endoglucanases which can be eff ective at broad pH with high activity.…”
Section: Discussionmentioning
confidence: 99%
“…GH12 endoglucanases usually presented low resistance to high pH, for example, cel12B (GH12) from Pectobacterium carotovorum subsp. carotovorum (Pcc) showed a pH optimum of 6.0 (Ibrahim et al, 2017), AcCel12B (GH12) from Acidothermus cellulolyticus 11B remained less than 10% of max enzyme activity at pH 7.0 (Wang et al, 2015). Especially, the neutral endoglucanase EgH31 showed outstanding stability in wide pH ranging from 3.0 to 11.0, and remained more than 70% of peak activity between pH 6.0 and 9.0, while most endoglucanases reported did not functionally work under the wide range of pH (Galante et al, 1998;Bhat, 2000;Trivedi et al, 2011;Yang et al, 2016;Zeng et al, 2016).…”
Section: Discussionmentioning
confidence: 99%
“…Recombinants cel12B , cel8C , β-glu , and peh28 encoding the respective cellulases, cel12B, cel8C, β-glucosidase, and polygalacturonase (peh28) expressed in E. coli were used for investigating their combined activities (cocktail formulation given below) on a grapefruit processing waste using their partially purified forms previously described …”
Section: Experimental Sectionmentioning
confidence: 99%
“…87128, C104450, and C0378, respectively) was added to all reactions each at 0.1 mg/g solid biomass. Hydrolysate samples (1.0 mL) were diluted with four volumes of ethanol and the filtrates containing mono- and disaccharide hydrolytic products were evaporated for derivatization and GC-MS quantification, as previously described . The GC (Model 6890, Agilent Technologies, Inc. Hewlett-Packard, Santa Clara, CA) operating conditions were as follows: 180 °C oven temperature for 2 min; increased by 230 °C for 15 min at 15 °C/min, 250 °C injector temperature, 1:10 split ratio, 1.0 μL injection sample/standard and 2 °C/min rate flow helium.…”
Section: Experimental Sectionmentioning
confidence: 99%